Boos, Julien, Kirk, David William, Dattler, Andre, Piccolotto, Mari-Luz, Zuercher, Werner, Gfeller, Sandro, Neuner, Philippe, Christensen, Jesper, Wishart, William L., Beverly, Michael, Von Arx, Fabian, Litherland, Karine, Van De Kerkhof, Esther, Swart, Pieter Jacob, Faller, Thomas, Beyerbach, Armin, Morrissey, David, Hunziker, Juerg and Beuvink, Iwan (2012) Whole-body scanning PCR; a sensitive method to study the biodistribution of mRNAs, noncoding RNAs and therapeutic oligonucleotides. Nucleic Acids Research, Nuclei. ISSN 0305-1048

Abstract

Efficient tissue-specific delivery is a crucial factor in the successful development of therapeutic oligonucleotides. Screening for novel delivery methods with unique tissue-homing properties requires a rapid, sensitive, flexible and unbiased technique able to visualize the in vivo biodistribution of these oligonucleotides. Here we present whole body scanning PCR, a platform that relies on the local extraction of tissues from a mouse whole body section followed by the conversion of target-specific qPCR signals into an image. This platform was designed to be compatible with a novel RTqPCR assay for the detection of siRNAs and with an assay suitable for the detection of heavily chemically modified oligonucleotides which we termed Chemical-Ligation qPCR (CL-qPCR). In addition to this, the platform can also be used to investigate the global expression of endogenous mRNAs and non-coding RNAs. Incorporation of other detection systems, such as aptamers, could even further expand the use of this technology.

Item Type:	Article
Related URLs:	http://www.ncbi.nlm.nih.gov/pubmed/23766...
Keywords:	siRNA,miRNA,mRNA,noncodingRNA,RTqPCR,Chemical-Ligation,Ligation,qPCR,whole body scanning PCR,RNA extraction
Related URLs:	http://www.ncbi.nlm.nih.gov/pubmed/23766...
Date Deposited:	13 Oct 2015 13:14
Last Modified:	13 Oct 2015 13:14
URI:	https://oak.novartis.com/id/eprint/7984